Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Western blot [3]
- Other assay [1]
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- Product number
- 14-1028-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Bcl-2 Monoclonal Antibody (Bcl-2/100), eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: This bcl-2/100 monoclonal antibody reacts to human Bcl-2 (B-cell Lymphoma 2), the founding member of a family of pro-apoptotic proteins which includes Bcl-XL, Bcl-w, and Mcl-1. Bcl-2 plays a role in protecting against apoptosis-inducing insults such as dexamethasone and staurosporine, as well as irradiation and cytokine withdrawal. The mechanism is thought to include regulation of mitochondrial membrane permeability, resulting in sequestration of caspase proforms, or by preventing the release of apoptosis-mediating mitochondrial factors, such as cytochrome c and AIF (apoptosis-inducing factor), into the cytoplasm. Bcl-2 is localized to the cytoplasmic side of the mitochondrial membrane, the nuclear envelope, and the endoplasmic reticulum. Interestingly Bcl-2 does not play a role in preventing cell death of autoreactive T cells in the thymus or Fas-mediated apoptosis. Two isoforms, termed alpha and beta of Bcl-2 have been identified and are a result of alternative splicing at the C-terminus. This antibody recognizes both forms.
- Antibody clone number
- Bcl-2/100
- Concentration
- 0.5 mg/mL
Submitted references Effects of hydrogen-occluding-silica microparticles on wound repair and cell migratory behavior of normal human esophageal epitheliocytes.
Influence of hydrogen-occluding-silica on migration and apoptosis in human esophageal cells in vitro.
Li Q, Tanaka Y, Miwa N
Medical gas research 2018 Apr-Jun;8(2):57-63
Medical gas research 2018 Apr-Jun;8(2):57-63
Influence of hydrogen-occluding-silica on migration and apoptosis in human esophageal cells in vitro.
Li Q, Tanaka Y, Miwa N
Medical gas research 2017 Apr-Jun;7(2):76-85
Medical gas research 2017 Apr-Jun;7(2):76-85
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Bcl-2 was achieved by transfecting HeLa cells with Bcl-2 specific validated siRNAs (Silencer® select Product # s194310, s1915). Western blot analysis (Fig. a) was performed using whole cell extracts from the Bcl-2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Bcl-2 Monoclonal Antibody (Product # 14-1028-82, 1 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Bcl-2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), Raji (Lane 2), T-47D (Lane 3), MDA-MB-231 (Lane 4), HeLa (Lane 5) and Jurkat (Lane 6). The blot was probed with Anti-Bcl-2 Monoclonal Antibody (Product # 14-1028-82, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 27 kDa band corresponding to Bcl-2 was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of BCL2 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042) (Assay ID CRISPR946648_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of BCL2 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa CAS9 (Lane 2), HeLa BCL2 KO (Lane 3) whole cell extracts. The blot was probed with Anti-Bcl-2 Monoclonal Antibody (Bcl-2/100), eBioscience™ (Product # 14-1028-82) using 1 µg/mL dilution and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to BCL2.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 Western blot analysis of activated caspase-3 and Bax/Bcl-2 ratio in KYSE-70 cells and HEEpiCs. Note: (A, B) The immuno-detected bands of activated caspase-3 (15 kDa), Bax (19 kDa), Bcl-2 (28 kDa), beta-actin (42 kDa) and alpha-tubulin (48 kDa) at different concentrations of H 2 -silica in both cell lines. (C, D) The ratios of band densities for Bax/Bcl-2 of KYSE-70 cells and HEEpiCs, respectively. (E, F) The ratios (%) of band densities for activated caspase-3 versus alpha-tubulin and for Bax/Bcl-2 of KYSE-70 cells and HEEpiCs, respectively. HEEpiCs: Human esophageal epithelial cells; H 2 -silica: hydrogen-occluding-silica.